JPS58116415A - Using method of cholesterol reducing agent - Google Patents

Abstract

PURPOSE:A cholesterol reducing agent, prepared by adding a mixture of vegetable sterol with a compound selected from a triterpenic alcohol to an animal fat or oil, and capable of exhibiting a very great reducing effect on the cholesterol in a small amount without causing problems in storing food nor taste. CONSTITUTION:A mixture of vegetable sterol contained in soybean seeds, coconuts or cotton seeds, with a triterpenic alcohol, e.g. cycloartenol, 2,4-methylenecycloartanol and cyclolaudenol, is added to an animal fat or oil or a food containing it, e.g. mayonnaise or dressing, etc. The triterpenic alcohol is contained in rice bran, soybean oil, etc., and commercially available gamma-oryzanol is saponified conveniently to prepare the aimed triterpenic alcohol from the unsaponified compound. The mixing ratio of the vegetable sterol to the triterpenic alcohol in the mixture is as follows: 100pts.wt. vegetable sterol and 1-20pts.wt., preferably about 5pts.wt., triterpenic alcohol.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method of using a cholesterol-lowering agent, and more particularly, it utilizes the synergistic effect of plant sterols and chitriterpene alcohols such as cycloartenol to reduce serum cholesterol levels. It provides animal fat and oil foods.

Cholesterol is known to play an important physiological role in the body, but on the other hand, it is said that when a large amount of cholesterol accumulates in the body, arteriosclerosis is likely to occur. For this reason, many cholesterol-lowering agents have been reported so far, including cholesterol synthesis inhibitors such as phenylethyl acetate os 7 It has been known. However, the above-mentioned cholesterol-lowering agents have drawbacks such as causing various side effects when used for a long period of time, impairing the flavor of the food when added to food, or having to be mixed in large quantities because of their low lowering effect.

The present invention has been made in view of the above circumstances, and has been researched for many years to obtain a cholesterol-lowering agent that does not change the taste of the food even when added to food, and has a cholesterol-lowering effect in a small amount. announced that there is a substance in the unsaponifiable matter of soybean oil that promotes the cholesterol-lowering effect of plant sterols (Kanen et al.; Nutrition and Food 19, 43).
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(1967). However, it was found that this lowering method had various drawbacks when applied directly to food production, and further research revealed that triterpene alcohols such as cycloartenol have a lower cholesterol-lowering effect of plant sterols. They discovered that plant sterols and triterpene alcohols such as 7-chloroartenol, 2,4-methylenecycloartanol, and cyclolaudenol were included in foods containing animal fats and oils to create a canine cholesterol-lowering agent with a lowering effect. The problem was solved by making the food containable.

The plant sterols used in the present invention are widely distributed in nature and have been known to have a cholesterol-lowering effect. It is known that it is distributed in, for example, soybean seeds, coconut seeds, cotton seeds, sesame seeds, sunflower seeds, and the like. To produce plant sterols from these seeds, for example, oils and fats are extracted or squeezed from the seeds, saponified by adding an alkali such as caustic soda or caustic potash, and the unsaponifiables are converted to an organic solvent such as ether, n- It can be produced by extracting it with xane or the like, distilling it to dryness, then dissolving it in methanol and cooling it, separating it and purifying it. Normally, the plant sterols obtained in this way contain many nosterols such as β-sitosterol, stigmasterol, and campesterol, and the types and contents of these substances vary depending on the type of raw material seeds, differences in extraction methods, etc. However, in the present invention, any substance having the effect of reducing serum sterols can be used, regardless of whether it is pure or impure.

Next, it is well known that triterpene alcohols such as cycloartenol, 2,4-methylenecycloartanol, or cyclolaudenol, which are mixed with the above plant sterols, exist in rice bran, but in addition to soybean oil, Contained in olive oil, corn oil, etc. The manufacturing method is, for example, to extract the oil from rice bran, saponify it with an alkali such as caustic soda or caustic potassium, extract the unsaponifiables with ether or other organic solvent, collect only the γ-oryzanol fraction, and add it to alcoholic caustic solution. Although it is produced by saponifying it with an alkali, extracting it with an organic solvent after saponification, and crystallizing it, it is convenient to saponify commercially available r-oryzanol and produce it from the unsaponified product. The cycloartenol etc. obtained in this way can usually have a purity of 80 to 99% by gas chromatography, but the present invention is not limited to such high purity. Crude products containing 20% or more can also be used. According to the research of the present inventors, the triterpene alcohol such as cycloartenol, 2,4-methylenecycloartanol or cyclolaudenol,
If plant sterols are lacking, no effect on lowering serum cholesterol will be exhibited unless one or more of them are added to the feed, but when used in combination with the above-mentioned plant sterols, a small amount will have a significant lowering effect. Such a synergistic effect was made possible by the present inventor's announcement, and is a completely new discovery.

Now, the synergistic effect will be explained by an experimental example using cycloartenol as an example.

The experiment was carried out by mixing plant sterols and cycloartenol in butter-containing feed, which has been shown to have a clear effect on raising serum cholesterol, through a feeding test using white rats. The plant sterols used at this time were soybean sterols produced from unsaponifiables of soybean oil, including approximately 50% β-sitosterol, stake mastirol, and canhesterol.
-#f Each contains about 25 units and has a cholesterol-lowering effect. Cycloartenol was produced by purifying γ-oryzanol with an organic solvent and saponifying the purified product, and its purity was 97.5% as determined by gas chromatography. The feed was prepared using a basic diet consisting of casein, P paper powder, inorganic salts, vitamins, and chlorinated purine mixed with butter (commercially available), which has the effect of raising serum cholesterol, and margari (commercially available), which is said to have no raising effect on serum cholesterol. ), with or without the addition of the soy sterol and cycloartenol, respectively,
The blending ratio was adjusted by adding sucrose, and the blending ratio is as shown in Table 1. However, the total amount is 100 copies.

Table 1 The animal test involved five groups of white rats (10 mice per group), the first group receiving butter, the second group receiving butter and soy sterols, and the third group receiving butter, soy sterols and cycloartenol. The fourth group was given margarine, and the fifth group was given margarine, soy sterol, and cycloartenol, and allowed to eat freely for 20 days. After 20 days, the serum was centrifuged, and the amount of cholesterol contained therein was determined by a colorimetric method using O-7 taraldehyde (manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.) and perchloric acid.

The above results are shown in Table 2. However, the right column of Table 2 is based on 15 parts of the butter in Table 1.

Table 2 As is clear from Table 2, total cholesterol content is extremely high in butter alone (group 1), but when soybean sterol is added to it (group ■), it shows a slight decrease; The present invention (■
ward) shows a significant decrease in total cholesterol. This phenomenon is also observed in margarine, which is said to have low cholesterol accumulation, and it is not a phenomenon that only occurs when animal fats and oils are fed, such as butter, but it is also due to the extremely large synergistic effect of the combination of plant sterols and cycloartenol. The purpose is to prove it.

Next, we compared the basic food with increased cholesterol and the addition of soy sterol alone, and the addition of a mixture of soy sterol and cycloartenol or 2,4-methylenecycloartanol. It was determined from the abundance ratio of plant sterol, cycloartenol, and 2,4-methylenecycloartanol in the unsaponifiable matter of soybean oil. That is, cycloartenol contained approximately 1/20 of the sterol in soybean oil. Therefore, we conducted an animal test in which feed was fed at this rate, and the results shown in Table 3 were obtained.

Table 3 According to the results, cycloartenol has a remarkable synergistic effect on the plasma cholesterol-reducing effect of soybean sterol, and 2,4-methylenecycloartanol also has a considerably strong synergistic effect, although the degree is different. . In another experiment, cycloartenol also showed a synergistic effect comparable to that of 2,4-methylenecycloartanol.

The cholesterol-lowering agent of the present invention is a mixture of a plant sterol and a triterpene alcohol as in the above example, and takes advantage of their synergistic effect. A synergistic effect can be observed at any mixing ratio with triterpene alcohols such as nol, but preferably 1 to 20 parts of the triterpene alcohol, especially the latter around 5 parts, is commonly used per 100 parts of plant sterol. The obtained preparation shows an extremely large reduction effect when mixed in a small amount, and even when added to food, it does not change the taste of the food and there is no problem in preserving the food, so it is especially suitable for mixing with animal fats and oils. , which has the effect of preventing the increase in cholesterol, which is a disadvantage of this. As the animal fats and oils, butter, lard, mutton fat, whale oil, etc., or foods containing these in large amounts, which are known to cause cholesterol accumulation in the body, are used. The amount added naturally varies depending on the type and content of animal fats and oils, but when added to butter, plant sterols 41 to 10t1 cycloartenol, 2,4
- Triterpene alcohol such as methylene cycloartanol or cyclolaudenol 200 m9 to 500 mIj
It is preferable that Additions can be made in solid form by kneading, or plant sterols and cycloartenol can be pre-contained in large quantities in edible oil or animal fat, and then added to foods. It may be added. The food obtained in this way lowers serum cholesterol in the human body and is therefore effective in preventing arteriosclerosis. Therefore, there is no need to take precautions such as refraining from ingesting butter or the like due to dietary restrictions, so there is an advantage that meals can be enjoyed.

The present invention will be explained in detail below with reference to Examples.

Example 1 1 kg of soybean oil was saponified with io% caustic soda, unsaponifiables were extracted with ether, and the extract was purified by methanol treatment to produce soybean sterols 3,5 containing 70% of total sterols.
I got t. This soybean sterol contained approximately 50% β-sitosterol, approximately 25% stigmasterol, and approximately 25% campesterol.

Separately, cycloartenol is γ prepared from rice bran.
- Oryzanol 10F in methanol and ethyl arsenate:
After repeated recrystallization with a mixture of ethanol (1:1), it was saponified with alcoholic potassium hydroxide and extracted with hexane. The purity of the obtained cycloartenol was 975% as determined by gas chromatography.

The above soybean sterol 680■ and cycloartenol 24
M9 was mixed with 340~/100t butter 1002 containing cholesterol, and mixed with Wistar male rats (100% in each group).
When the rats were fed ad libitum and reared for 30 days, the serum cholesterol level was 11.
The cholesterol-lowering effect was 130m97dl for the soybean sterol and cycloartenol-deficient cholesterol-lowering effect.

Furthermore, when butter mixed with the formulation of the present invention was tested for taste by a panel of 10 people, no significant difference was observed between butter mixed with the formulation of the present invention and butter not added.

Example 2 Sesame oil collected from sesame seeds was treated in the same manner as in Example 1 to obtain 3.8f of sesame sterols. This was mixed with cycloartenol produced by the method shown in Example 1, mixed with mutton tallow in the same manner as in Example 1, and the same animal test as in Example 1 was conducted. As a result, 9 significant cholesterol reduction results were obtained. showed that.

Example 3 Soybean sterol was prepared in the same manner as in Example 1, and 2゜4-methylenecycloartanol was extracted with hexane after saponifying γ-oryzanol, distilling off hexane from the resulting unsaponifiable matter, and extracting with methanol. It was crystallized in Obtained 2,
The purity of 4-methylenecycloartanol is 83.5%
Met. This substance 24rn9 and the soybean sterol 6
The mixture with 801n9 was mixed with 1 oor of the butter of Example 1, and an animal test was conducted using the method of Example 1. As a result, a remarkable cholesterol-lowering effect was observed.

Procedures Book dated March 1, 1981, Patent Section 11, No. 1, Public Notice of the Case, Patent Application No. 214243 of 1982, Name of the Invention, Method of Using Cholesterol-Lowering Agents, 3, Person Making the Amendment, Relationship with the Case, Patent Applicant 5: Number of inventions increased by supplementation Target of amendment: Specification 2 Correction of content of supplement 1: Title of invention: Method of using cholesterol-lowering agent 2: Claims (1) Plant sterol 1. A method of using a cholesterol-lowering agent characterized by mixing it with a selected compound such as tritylben alcohol and adding it to animal oil or fat or a food containing the same.

+2) A method for using the cholesterol-lowering agent according to item 1 of the scope of water within patent a, characterized in that the mixing ratio of 8m sterol dotrityl bene alcohol is 1 to 20 parts of the latter to 100 m of the former.

3.Details of the invention ii5! The present invention relates to a method of using a cholesterol-lowering agent, and more particularly, it provides an animal fat and oil food that reduces serum cholesterol levels by utilizing the synergistic effect of plant sterols and triterpene alcohols such as cycloartenol. There's something to do.

Cholesterol is known to play a variety of physiological roles in the body, but on the other hand, it is said that when large amounts of cholesterol accumulate in the body, arteriosclerosis is likely to occur. For this reason, more cholesterol-lowering drugs have been recommended than before9.
Known cholesterol synthesis inhibitors include phenylethyl acetate, phenylbutyrate, and triparanol; examples of natural lowering agents include linoleic acid, lecithin, and adrenal hormone. However, the above-mentioned cholesterol-lowering agents have drawbacks such as having excessive copper effects when used for a long period of time, and when added to food, they may harm the flavor of the food or have little lowering effect and must be mixed in large quantities. I'm worried.

The present invention was made in view of the above-mentioned circumstances.The present invention was developed after many years of research in order to obtain a cholesterol-lowering agent that does not change the taste of food even when added to food, and has a large cholesterol-lowering effect even in small amounts. It was announced that there is a substance in the unsaponifiable matter of soybean oil that promotes the cholesterol-lowering effect of plant sterols (Kinjirushi et al.; Nutrition and Food 19, 43).
9 members.

(1967)). However, it was found that this lowering method had many disadvantages when applied directly to food production, and as a result of extensive research funding, it was found that triterbene alcohol was It was discovered that plant sterols significantly enhance the cholesterol-lowering effect, and by incorporating plant sterols and triterpene alcohols such as cycloartenol, 2,4-methylenecycloartanol, and cycloartenol into animal fat-containing foods, the cholesterol-lowering effect was achieved. I decided to solve the problem by using foods that contain a great cholesterol-lowering agent.

The plant sterols used in the present invention are widely distributed in nature and have been known to have a cholesterol-lowering effect. It is known that it is distributed in, for example, soybean pickled plums, coconut seeds, cotton seeds, sesame seeds, sunflower seeds, and the like. The production of A-type sterols from these toothpicks involves, for example, extracting fats and oils from the above-mentioned toothpicks, adding alkali such as caustic soda and caustic potash to this, and saponifying the unsaponifiables with an organic solvent. For example, it can be produced by extracting it with ether, n-hexane, etc., distilling it to dryness, dissolving it in methanol, and cooling it. Many of the substances include β-sitosterol, stigmasterol, and campesterol9, and the types and contents of these substances vary depending on raw material seeds, type 011 extraction methods, etc. In the present invention, any substance having the effect of reducing serum sterol O can be used, regardless of whether it is pure, pure, or #1.

Next, it is well known that triterpene alcohols such as cycloartenol, 2,4-methylene Schiff n-artanol, or cycloartenol, which are mixed with the above-mentioned plant sterols, exist in rice bran, but in addition to soybean oil, olive oil, etc. , is stored in chili oil, etc. The manufacturing method is, for example, by extracting oil from rice bran.
Saponify it with an alkali such as potassium, extract the non-cannibalized substances with ether or other organic solvents, remove only the r-orizinol fraction, saponify it with an alcoholic alkali, and extract with an aS agent after saponification. It is produced by saponifying commercially available r-oryzanol or by crystallizing it, but it is convenient to produce it from the unsaponified product. The cycloartenol etc. obtained in this way can usually have a purity of 80 to 99 by gas chromatography, but the present invention is not limited to such purity. %2016 or more om or more can also be used. According to the research conducted by the present inventors, triterpene alcohols such as cycloartenol, 2,4-methylenecycloartanol, and cyclolaudenol should not be added to feeds containing more than 1 liter of phytosterols when they are lacking in plant sterols. Although it does not show a cholesterol-lowering effect,
When used in combination with the above-mentioned plant sterols, a small amount can produce a significant lowering effect. This synergistic effect has been announced by the present inventor and is a completely new discovery.

Now, the synergistic effect will be explained by an experimental example using cycloartenol as an example.

The experiment was carried out using a feeding test using white mice, in which plant sterols and cycloartenol were added to putter-containing feed, which had a clear effect on raising serum levels of restirol. The nine sterols used at this time are soybean sterols produced from unsaponifiable matter of soybean oil, and this contains about 45.91ks of cytostyrene. Sterol 26.7% and campesterol approximately 2 & 9L Putushika Styrol α
5. Contains Schiff, which has a cholesterol-lowering effect. Also, cycloartenol contains r-oryznol! Ml with solvent! ! L, purified product T saponified and manufactured, its cotton & Fi gas chromatography! The purity according to Togufi was 97.5-. Feed 111#! casein, 70% rose, inorganic salts, vitamins, chlorinated purine,
Cottonseed oil etc. are mixed to make Nine Basic Steel T-114, among which the cottonseed oil is changed to butter oil which has the effect of raising serum cholesterol, and the soybean sterol and cycloartenol are added or not, respectively. The blending ratio was adjusted by adding sucrose to the specific gravity tube, and the blending ratio was F181111.
! ! This is the circular shown in . However, the total amount is 100 copies.

Table 1: Animal testing of four groups of white junipers? (1 review lo) to glN
K uses basic feed, the second group uses putter oil instead of cottonseed oil, and the 4143 group uses butter oil! sterol,
The fourth animal was fed putter oil, soybean tail, and Schiff's artenol, and allowed to eat freely for 20 days. 2
0 days later W! centrifuged and the amount of cholesterol contained therein to-7 taraldehyde (manufactured by Wako #1s)
and perchlorine II! P was quantified by colorimetry using .

The above results are shown in Table 2.

Table 2) As can be seen, m* and hepatic cholesterol are extremely high in the area containing cottonseed oil (Archive 1), which has changed to tree oil (Archive ■), but with the addition of soybean sterols (Archive 1).
Section 11) shows a significant reduction in cholesterol. However,
When soybean sterol and cycloartenol were added (dish group), cholesprol, which was even stronger, had a negative effect, and the values were lower in the animals fed the basic diet.

Next, we produced a single meal that contained almost no cholesterol, increased the amount of cholesterol, added soy sterol alone, and added soy sterol and cycloartenol or 2,4-methylenecycloaltenol. The ratio of plant sterols and cycloartenol in the unsaponifiable matter of soybean oil compared with the mixture of tanol added is 2.
This was determined based on the abundance ratio of 4-methylenecycloartanol, and cycloartenol contained approximately 1/2 of the sterol in soybean oil. Therefore, when we conducted an animal test using this ratio of 151%, we obtained the results shown in Table 3.

According to the results of the third paper, cycloartenol has a significant synergistic effect on the plasma cholesterol-reducing effect of soybean sterol, and 2,4-methylene 7-chloroartanol also has i! [F1
Although they are different, they have a fairly strong synergistic effect. In another experiment, cyclolacdenol showed a synergistic effect comparable to that of 42.4-methylenecycloartanol.

The cholesterol-lowering agent of the present invention is made by mixing the seed sterol triterben alcohol as in the above example and utilizing the synergistic effect thereof, but it also contains plant sterol and cycloartenol, 2,4-methylenecycloartanol, or cycloartanol. Although a synergistic effect is observed at any mixing ratio with triterpene alcohol such as artenol, it is preferable to use 1 to 20 parts of triterpene alcohol, especially the latter 5 parts, per 100 parts of seed sterol. . The obtained preparation shows an extremely large reduction effect when mixed in a small amount, and even when added to food, it does not change the taste of the food and there is no problem in preserving the food, so it is especially suitable for mixing with animal fats and oils. However, it has the effect of preventing an increase in cholesterol, which eliminates its drawbacks. The above-mentioned animal fats and oils and foods include putter, lard, mutton tallow, whale oil, and foods containing these.In general, when it comes to money-making, etc., if the salad oil is vegetable-based, it is vegetable-based. Although it is classified as an oil and fat food, since the egg yolk used to emulsify salad oil and vinegar contains lipid (egg yolk oil), it is included in the animal oil Mkt-containing food in the present invention. The same applies when egg yolk is used in dressings; in short, it includes anything that causes cholesterol to accumulate in the body after ingestion.

It is obvious that the amount of plant sterols etc. to be added varies depending on the type and content of animal fats and oils, but when adding it to a putter, plant sterols 4F to 10% are added to the putter lh.
F, Schiff-attack altenol, 2° 4-methylene Schiff-attack altanol or cyclo2j'/-le 4Loto! The J terpene alcohol is preferably 200 to 5ooH9. Additions may be made by kneading in a planar form, and may include plant stems,
A large amount of cycloartenol may be pre-contained in edible oil or animal oil and fat, and this may be added to food by the method of /9Tii. The nine foods obtained in this way are effective in preventing arteriosclerosis by lowering serum cholesterol in the human body. Since there is no obligation to take care of the temple, one can gather food and make one's YftlA. .

Hereinafter, 111 aspects of the present invention will be explained with reference to Examples.

Actual IM Example 1 Soybean oil 1 was saponified with 9t 10% caustic soda, unsaponifiables were extracted with ether, the extract was purified by methanol treatment, and total sterols were 70$1i-containing soybean sterols 3
．． Obtained 5F. This soybean sterol contained about 50% of β-sitosterol, about 25% of stigmasterol, and about 25% of campesterol.

In addition to this, cycloartenol was prepared from rice bran.
- Oryzanol 10t in methanol and ethyl acetate:
After repeated NM with ethanol (1:1) fi [,
It was purified with alkaline potassium hydroxide and extracted with hexane. The purity of the obtained cycloartenol was determined to be 97.511 by gas chromatography.

The above soybean sterol 680M9 and cycloartenol 2
4Wvft:zLz, Xtea-/l/contains 34G! /1
00t putter 10Of and fed ad libitum to Quistar male rats (1θ animals for each evaluation) and kept for 30 days, the serum cholesterol level was lower than that of the mixture of the present invention added to sardine/ril15d. A130II, which lacks soybean styrene and cycloartenol, was rare in /a.
Cholesterol lowering effect was observed at g/hz.

Furthermore, when the formulation tm of the present invention was tested for taste on a panel of 110 putters, no significant difference was found between the formulation and the formulation without the addition.

Example 2 Sesame oil collected from sesame seeds was treated in the same manner as in Example 1 to obtain 8 tons of sesame sterules. This was mixed with cycloartenol produced by the method shown in Example 1, and Example 1 was prepared. It was mixed with mutton tallow in the same manner as in Example 1, and the same animal test as in Example 1 was conducted, and as a result, it showed a significant cholesterol-lowering result.

Example 3 Soybean sterols! li! Prepared in the same manner as in Example 1, 2°4-methylenecycloartanol was r-oryzanol t
After saponification, the mixture was extracted with hexane, and the hexane was distilled off from the resulting non-saponified product, which was then crystallized with methanol. The purity of the obtained 2,4-methylenecycloartanol is 8&5.
It was 11. this? A mixture of 241 v of soybean sterol and 680 da of the soybean sterol was mixed with 1 oor of the putter of Example 1, and an animal test was conducted according to the method of Example 1. As a result, a remarkable cholesterol-lowering effect was observed.

Example 4 Vinegar (iron [6%) 12F, mustard powder 3F, salt 2F%
? @1 f and Ulu yellow liquor 12 f (fil type 5
In the aqueous phase raw material 30f consisting of 0 Is), 30 Da of soybean sterol (approximately 0.7-2 Da prepared in the same manner as in Example 1 based on egg yolk oil) and 2 Da cycloartenol (approximately 2 Da relative to egg yolk oil) were added. α04Is: Adjusted in the same manner as in Example 1)t
- Added 70 ftm of soybean salad oil and mixed to produce YW naze.

patent applicant Food Research Foundation

Claims (2)

[Claims] (1) A method of using a cholesterol-lowering agent, which comprises adding a mixture of a plant sterol triterpene alcohol and a selected compound to animal fats and oils or foods containing the same. (2) The method of using the cholesterol-lowering agent according to claim 1, wherein the mixing ratio of the plant sterol triterpene alcohol is 1 to 20 parts of the latter to 100 parts of the former.